U.S. patent number 10,314,911 [Application Number 14/679,667] was granted by the patent office on 2019-06-11 for methods for protecting and treating traumatic brain injury, concussion and brain inflammation with intranasal insulin.
This patent grant is currently assigned to HealthPartners Research & Education, The Henry M. Jackson Foundation for Advancement of Military Medicine, Inc.. The grantee listed for this patent is HealthPartners Research & Education. Invention is credited to Fiona Brabazon, Leah Ranae Bresin Hanson, Kimberly Byrnes, William H. Frey, II.
United States Patent |
10,314,911 |
Frey, II , et al. |
June 11, 2019 |
Methods for protecting and treating traumatic brain injury,
concussion and brain inflammation with intranasal insulin
Abstract
The present system is directed in several embodiments to a
method of administration of a therapeutic composition for
protection of the brain of a subject at risk of injury leading to
traumatic brain injury (TBI) and/or treatment of injury to the
brain resulting from TBI. The method includes administering one or
more therapeutic compositions comprising an effective amount of
insulin directly to the subject patient's CNS, with no to minimal
systemic exposure. Preferably, this method comprises administration
of an effective amount of insulin to the upper third of a patient's
nasal cavity, thereby bypassing the patient's blood-brain barrier
and delivering the therapeutic composition directly to the
patient's central nervous system.
Inventors: |
Frey, II; William H. (St. Paul,
MN), Bresin Hanson; Leah Ranae (Vadnais Heights, MN),
Byrnes; Kimberly (Gaithersburg, MD), Brabazon; Fiona
(Silver Springs, MD) |
Applicant: |
Name |
City |
State |
Country |
Type |
HealthPartners Research & Education |
Minneapolis |
MN |
US |
|
|
Assignee: |
HealthPartners Research &
Education (Minneapolis, MN)
The Henry M. Jackson Foundation for Advancement of Military
Medicine, Inc. (Bethesda, MD)
|
Family
ID: |
54208773 |
Appl.
No.: |
14/679,667 |
Filed: |
April 6, 2015 |
Prior Publication Data
|
|
|
|
Document
Identifier |
Publication Date |
|
US 20150283065 A1 |
Oct 8, 2015 |
|
Related U.S. Patent Documents
|
|
|
|
|
|
|
Application
Number |
Filing Date |
Patent Number |
Issue Date |
|
|
61976634 |
Apr 8, 2014 |
|
|
|
|
Current U.S.
Class: |
1/1 |
Current CPC
Class: |
A61K
38/28 (20130101); A61K 45/06 (20130101); A61P
5/50 (20180101); A61K 38/28 (20130101); A61K
2300/00 (20130101); A61K 9/0043 (20130101) |
Current International
Class: |
A61K
38/28 (20060101); A61P 5/50 (20060101); A61K
45/06 (20060101); C07K 14/62 (20060101); A61K
9/00 (20060101) |
References Cited
[Referenced By]
U.S. Patent Documents
|
|
|
2989437 |
June 1961 |
Wruble et al. |
5135923 |
August 1992 |
Siren |
5624898 |
April 1997 |
Frey, II |
5849290 |
December 1998 |
Brown et al. |
5874573 |
February 1999 |
Winchell et al. |
5939395 |
August 1999 |
Yu et al. |
6113906 |
September 2000 |
Greenwald et al. |
6180603 |
January 2001 |
Frey, II |
6313093 |
November 2001 |
Frey, II |
6342478 |
January 2002 |
Frey, II |
6407061 |
June 2002 |
Frey, II |
6413499 |
July 2002 |
Clay |
6544542 |
April 2003 |
Sonoke et al. |
6576660 |
June 2003 |
Liao et al. |
7205276 |
April 2007 |
Boderke |
2001/0043915 |
November 2001 |
Frey, II |
2001/0047032 |
November 2001 |
Castillo et al. |
2002/0028786 |
March 2002 |
Frey, II et al. |
2002/0072498 |
June 2002 |
Frey, II |
2002/0082215 |
June 2002 |
Frey, II |
2002/0133877 |
September 2002 |
Kuiper et al. |
2002/0141971 |
October 2002 |
Frey, II |
2003/0072793 |
April 2003 |
Frey, II et al. |
2003/0165434 |
September 2003 |
Reinhard et al. |
2003/0229025 |
December 2003 |
Xiao et al. |
2004/0101521 |
May 2004 |
Andersen |
2008/0020975 |
January 2008 |
Wynick |
2008/0305077 |
December 2008 |
Frey, II |
2012/0322727 |
December 2012 |
Abdel Maksoud |
2013/0090317 |
April 2013 |
Vanlandingham et al. |
2014/0031280 |
January 2014 |
Frey, II |
|
Foreign Patent Documents
|
|
|
|
|
|
|
WO90/00057 |
|
Jan 1990 |
|
WO |
|
WO91/07947 |
|
Jun 1991 |
|
WO |
|
WO98/42275 |
|
Oct 1998 |
|
WO |
|
WO08/62420 |
|
May 2008 |
|
WO |
|
Other References
Anonymous. "Severe TBI Symptoms"
www.traumaticbraininjury.com/symptoms-of-tbi/severe-tbi-symptoms/.
Published Jul. 27, 2012. cited by examiner .
Anonymous. "Mild TBI Symptoms"
www.traumaticbraininjury.com/symptoms-of-tbi/mild-tbi-symptoms/.
Published Aug. 18, 2016. cited by examiner .
Anonymous. "Diagnosis"
www.traumaticbraininjury.com/symptoms-of-tbi/diagnosis/. Published
Jul. 27, 2012. cited by examiner .
Ley et al. "Diabetic Patients With Traumatic Brain Injury: Insulin
Deficiency Is Associated With Increased Mortality" J. of Trauma 70:
1141-1144. (Year: 2011). cited by examiner .
Meierhans et al. "Brain metabolism is significantly impaired at
blood glucose below 6 mM and brain glucose below 1 mM in patients
with severe traumatic brain injury" Critical Care 14:R13 (Year:
2010). cited by examiner .
Cerecedo-Lopez et al. "Insulin-associated neuroinflammatory
pathways as therapeutic targets for traumatic brain injury" Medical
Hypotheses 82:171-174. (Year: 2013). cited by examiner .
Brabazon et al. "Intranasal Insulin Treatment of Traumatic Brain
Injury" J. Neurotrauma 31:A-106, Abstract D1-17. (Year: 2014).
cited by examiner .
International Preliminary Report on Patentability dated Oct. 20,
2016 for International Application No. PCT/US2015/024621 filed Apr.
7, 2015. cited by applicant .
Dezhi et al., "HIF1 alpha upregulation and neuroprotection with
deferoxamine in a rat neonatal stroke model" Pediatric Research,
55(4): 408A (Apr. 2004). cited by applicant .
Ross et al., Intranasal administration of interferon beta bypasses
the blood-brain to target the central nervous system and cervical
lymph nodes: a non-invasive treatment strategy for multiple
sclerosis, Journal of Neuroimmunology, 151(1-2): 66-67 (Jun. 2004).
cited by applicant .
Adachi et al (Brit J Rheumatol 36:255-259, 1997). cited by
applicant .
Jarvinen K and Uritti A. Duration and long-term efficacy of
phenylephrine-induced reduction in the systemic absorption of
ophthalmic timolol in rabbits. J Ocul. Pharmacol. 1992; 8(2):91-98;
abstract only. cited by applicant .
Vachharajani NN et al. A pharmacokinetic interactioni study between
butorphanol and sumatriptan nasal sprays in healthy subjects:
importance of the timing of butorphanol administration.
Cephalalgia, 2002; 22:282-287. cited by applicant .
Kruck et al., Clin Pharmacol Ther, 48(4): 439-446, Oct. 1990. cited
by applicant .
Gordon et al., Amer J Med Sci, 297(5): 280-284, May 1989. cited by
applicant .
Wang and Semenza, Blood, 82(12): 3610-3615, Dec. 15, 1993. cited by
applicant .
P. Murali Doraiswamy and Anne E. Finefrock, Metals in our minds:
therapeutic implications for neurodegenerative disorders, The
Lancet Neurology vol. 3, Jul. 2004 (pp. 431-434). cited by
applicant .
Maxwell and Salniknow, Cancer Biology and Therapy 3(1): 29-35.
(Jan. 2004). cited by applicant .
Brenneisen et al., The Journal of Biological Chemistry 273(9):
5279-5287. (Feb. 27, 1998). cited by applicant .
Crapper McLachlan et al., Lancet 337(8753): 1304-1308. (Jun. 1,
1991). cited by applicant .
Chaston and Richardson, American Journal of Hematology 73: 200-210
(2003). cited by applicant .
King RG, Med J Aust, 142(6; 352, Mar. 18, 1985). cited by applicant
.
Youdin et al., Ann NY Acad Sci, 1012:306-325, Mar. 2004. cited by
applicant .
Lan and Jiang, J Neural Transmission, 104:469-481, 1997. cited by
applicant .
S. Talegaonkar, P.A. Mishra, Intranasal delivery: An approach to
bypass the blood brain barrier, Indian J. Phermacol, Jun. 2004,
vol. 36, Issue 3 140-147. cited by applicant .
Gould et al., "Glycogen Synthase Kinase-3: A Target for Novel
Bipolar Disorder Treatment," Jan. 31, 2004 (Jan. 31, 2004). The
Journal of Clinical Psychiatry, vol. 65, Is. 1; p. 1021; especially
abstract; p. 13, col. 2, para 3; p. 15, col. 1, para 4 to col. 2,
para 1; p. 17, col. 1, para 2. cited by applicant .
The Merck Index, Twelfth Edition, 1996, entries 3908 and 7135.
cited by applicant .
Venters Jr., Homer D. et al., "Heme from Alzheimer's brain inhibits
muscarinic receptor binding via thiyl radical generation" Brain
Research, 1997, 764, 93-100. cited by applicant .
Kornberg, Arthur, et al.; "Inorganic Polyphosphate: A Molecule of
Many Functions"; Annual Review Biochemistry, vol. 68: 89-125;
Annual Reviews; US 1999. cited by applicant .
Frey II, William H. et al.,; "Brain Research 714 (1996) 87-94:
Endogenous Alzheimer's brain factor and oxidaized glutathione
inhibit antagonist binding to the muscarinic receptor"; Elsevier
Science B.V.; US 1996. cited by applicant .
Frey II, William H. et al., "Brain Research 655 (1994) 153-160:
Inhibitor of antagonist binding to the muscarinic receptor is
elevated in Alzheimer's brain"; Elsevier Science B.V.; US 1994.
cited by applicant .
Otterbein, Leo E., et al.; "Invited Review: Heme Oxygenase: colors
of defense against cellular stress"; The American Physiological
Society; www.aiplung.orq: US2000. cited by applicant .
Rogers et al (Arch Intern Med 158:1021-1031, 1998). cited by
applicant .
van Beek et al (Biochem Biophys Res Comm 255:491-494, 1999). cited
by applicant .
Pahan et al (J Clin Invest 100:2671-2679, 1997). cited by applicant
.
Zhao et al (J Neurosci Res 52:7-16, 1998). cited by applicant .
Fawcett et al (Brain Res 950:10-20, 2002). cited by applicant .
Atack et al (J Neurochem 60:652-658, 1993). cited by applicant
.
Liu et al., Molecular and Cellular Biology, Sep. 1992, 3978-3990.
cited by applicant .
Frey et al., "Delivery of 1251-NGF to the Brain via the Olfactory
Route", Drug Delivery, 4:87-92, 1997. cited by applicant .
Ostovic et al (Pharm Res 10:470-472, 1993). cited by applicant
.
Rooijen (Calcif Tissue Int 52:407-410, 1993). cited by applicant
.
Body et al (Annals of Oncology, 5:359-363, 1994; Abstract Only).
cited by applicant .
Backstrom et al (J Neurosci 16:7910-7919, 1996). cited by applicant
.
Brabazon, F., et al., "Intranasal insulin treatment of traumatic
brain injury", (Jun. 1, 2014), XP055421658,
URL:https://tbitherapy.com/wp-content/uploads/2016/06/neurotrauma-poster--
2014-2.pdf. cited by applicant .
Brynes, Kimberly R., et al., "FDG-PET imaging in mild traumatic
brain injury: a critical review", Frontiers in Neuroenergetics,
vol. 5, Jan. 1, 2014 (Jan. 1, 2014), XP055421662, ISSN: 1662-6427,
DOI: 10.3389/fnene.2013.00013. cited by applicant .
Schilling,Thomas M., et al., "Intranasal insulin increases regional
cerebral blood flow in the insular cortex in men independently of
cortisol manipulation", Human Brain Mapping, vol. 35. No. 5, Aug.
1, 2013, pp. 1944-1956, XP055421678, ISSN: 1065-9471, DOI:
10.1002/hbm22304. cited by applicant .
Extended Search Report dated Nov. 29, 2017 issued by European
Patent Office for related application No. 15776436.6. cited by
applicant.
|
Primary Examiner: Miknis; Zachary J
Attorney, Agent or Firm: Barnes & Thornburg LLP Stone;
Jeffrey R.
Parent Case Text
CROSS-REFERENCE TO RELATED APPLICATIONS
This application claims priority to App. Ser. No. 61/976,634,
entitled "Method of Treating and/or Preventing Injury to the Brain
Caused by Traumatic Brain Injury by Intranasal Administration of
Insulin," filed Apr. 8, 2014, the entire contents of which are
hereby incorporated by reference in its entirety.
Claims
We claim:
1. A method for treating traumatic brain injury (TBI) in a patient,
comprising: administering a therapeutic agent consisting of at
least an effective amount of insulin to the upper third of the
nasal cavity of the patient, and thereby enabling at least an
effective amount of insulin to directly access the patient's
central nervous system by bypassing the blood-brain barrier; and
treating the patient's TBI by protecting the patient's brain from
inflammation and reducing inflammation in the patient's brain by
increasing the expression of anti-inflammatory M2 microglia cells
in the patient's brain.
2. The method of claim 1, further comprising administering the
insulin to a tissue innervated by the olfactory nerve, wherein the
administered insulin bypasses the blood-brain barrier to access the
patient's central nervous system to treat the TBI.
3. The method of claim 2, further comprising the administered
insulin bypassing the blood-brain barrier by migrating along a
neural pathway into the patient's central nervous system to treat
the patient's TBI.
4. The method of claim 1, wherein the administered insulin
comprises a non-zinc insulin.
5. The method of claim 1, further comprising pretreating the
patient's nasal cavity with an effective amount of at least one
vasoconstrictor before administering the insulin to the upper third
of the patient's nasal cavity.
6. The method of claim 1, wherein the at least an effective amount
of insulin is in the range of 1.times.10.sup.-7 to 0.1 mg/kg, with
reference to the patient's body weight.
7. The method of claim 6, wherein the range for the at least an
effective amount of insulin is 1.times.10.sup.-4 to 0.1 mg/kg, with
reference to the patient's body weight.
8. The method of claim 1, wherein the concentration of insulin in
the brain of the patient after a single dose is in the range of
1.times.10.sup.-13 to 1.times.10.sup.-9 molar.
Description
FEDERAL FUNDING
None
BACKGROUND OF THE INVENTION
Field of the Invention
The present invention is directed to methods of protection of the
brain of a subject at risk of injury leading to traumatic brain
injury (TBI) and/or treating injury to the brain in patients with
TBI. More particularly, the present disclosure is directed to a
method of protection of the brain of a subject at risk of injury
leading to TBI and/or treating TBI by administration of an
effective amount of insulin to the upper third of the patient's
nasal cavity.
Description of the Related Art
Traumatic Brain Injury (TBI) occurs when sudden trauma causes
damage to the patient's brain, e.g., when the head is suddenly and
violently hit by an object or when an object pierces the skull and
enters the patient's brain tissue.
The most common causes and risk activities for TBI include falls,
vehicle crashes and sports injuries. Indirect forces that jolt the
brain violently within the skull, e.g., shock waves from a
battlefield explosion also may result in TBI as may bullet wounds
or other brain-penetrating injuries.
Symptoms of a TBI include unconsciousness, inability to recall the
traumatic event, confusion, headache--including a headache that
will not go away or worsens with time, lightheadedness, dizziness,
blurred vision or tired eyes, ringing in the ears, bad taste in the
mouth, fatigue, lethargy, a change in sleep patterns, behavioral or
mood changes, trouble with memory, concentration, attention or
thinking, repeated vomiting, nausea, convulsions, seizures, an
inability to awaken from sleep, dilation of one or both pupils of
the eyes, trouble speaking coherently--including slurring of
speech, weakness or numbness in the extremities, unsteadiness, lack
of coordination, restlessness and agitation.
TBI is a threat to an individual's health in at least the following
ways: (1) TBI has direct effects, a short listing of these are
provided above; (2) Certain TBI's may increase the risk of
developing Alzheimer's disease and certain forms of dementia; and
(3) Repeated TBI, such as those that can occur in contact sports
such as football, boxing, hockey, lacrosse and soccer to name a
few, maybe linked to an increased risk of a dementia known as
chronic traumatic encephalopathy.
As defined herein, TBI, in addition to the above, includes
concussion injuries; concussions being a type of TBI.
Thus, patients at risk, as referred to herein, may comprise
individuals engaged in contact sports who are at risk for head
injuries as well as those individuals in professions, e.g.,
soldiers, police officers, fire fighters, athletes in contact
sports and the like, that places them at risk of head injuries
leading to TBI.
So far as we are aware, the only preventive, or protective,
treatment currently available includes protective gear such as
helmets for patients at risk of brain injuries that may lead to
TBI. Once TBI is diagnosed, the primary focus and treatment
comprises ensuring the patient's brain is properly oxygenated, with
sufficient blood flow and control of blood pressure. More severe
cases may require treatment involving physical therapy,
occupational therapy, speech and language therapy, physical
medicine, and psychological and/or psychiatric therapy.
Delivery of the agent and/or composition to the upper one third of
the patient's nasal cavity is a means of bypassing the BBB to
administer therapeutic compounds and/or agents directly to the CNS.
Evidence exists that intranasal treatment with certain therapeutic
agent(s) improves, i.e., prevents, protects against and/or treats,
a variety of neurological and psychiatric disorders, e.g., stroke,
in animals. This basic methodology is discussed and described in
U.S. Pat. No. 5,624,898 to Frey II entitled Method for
Administering Neurologic Agents to the Brain, as well as in U.S.
Pat. No. 6,313,093 to Frey II, the entire contents of each of which
are hereby incorporated by reference. This administration technique
is a vast improvement over systemic administration methods such as
intravenous and oral administration of drugs which generally cannot
cross the BBB to reach their targets within the CNS. In addition,
Frey's intranasal method is a significant improvement over the
general inhalation methods which target the lower two-thirds of the
patient's nasal cavity. Both the systemic and general intranasal
method targeting the lower two-thirds of the nasal cavity result in
a very large, unwanted and potentially dangerous systemic exposure
to the administered drug or therapeutic agent(s). The present
invention addresses, inter alia, this general intranasal problem as
well as ensures that the patient's non-CNS, systemic disease and/or
condition is protected from exposure to the therapeutic agent
administered to the upper third of the nasal cavity, and potential
harm therefrom.
General inhalation methods to the lower two-thirds of the nasal
cavity delivered by, e.g., nasal spray bottles, on the other hand,
result in a large amount of systemic absorption and exposure, with
a very small amount of the administered compound, i.e., less than
5%, making the tortuous journey around the turbinates to the upper
third of the nasal cavity and still less compound than that very
small amount further bypassing the BBB to actually reach the
CNS.
Delivery and administration to the upper third of the nasal cavity,
is very effective in administering the subject compounds or agents
to the desired target, i.e., the CNS, without significant systemic
exposure, though some systemic exposure does occur as is further
discussed below.
Unwanted systemic exposure of therapeutics used to treat CNS
diseases create several serious problems. The systemic metabolism
greatly reduces the bioavailability of any agent and/or compound
exposed to the non-CNS system. This reduction of bioavailability is
increased by unwanted plasma protein binding of the agent and/or
compound. As a result, only a small amount of the active
therapeutic agent and/or compound actually reaches the CNS. Because
of these, inter alia, issues, the actual dose that must be
administered in order to achieve a therapeutic dose in the targeted
CNS is far larger than the therapeutic dosing. As a consequence, a
relatively large concentration of the agent(s) and/or compounds(s)
is in the system and will affect non-target systemic organs and
systems. This can create unwanted and often dangerous side effects
on these non-target organs and systems, particularly in the
specific case of patient's having a systemic, non-CNS disorder or
condition that contraindicates the systemic use or exposure of the
therapeutic agent(s) needed to treat a CNS-related disorder or
condition.
We have addressed the efficiency needs in patent application Ser.
No. 12/134,385 to Frey II, et al., entitled "Pharmaceutical
Compositions and Methods for Enhancing Targeting of Therapeutic
Compounds to the Central Nervous System, the entire contents of
which are hereby incorporated by reference, and wherein a
vasoconstrictor is administered to the patient's nasal cavity
either just prior to, or in combination with, administration of at
least one therapeutic agent and/or pharmaceutical composition(s)
comprising a therapeutic compound(s) and/or agent(s). The
efficiency of the direct administration of the pharmaceutical
compound to the CNS, with concomitant reduction of systemic
exposure of the pharmaceutical compound is remarkable.
Moreover, we provide disclosure of the following patents and
applications, each of which are commonly assigned with the present
application and incorporated herein in their entirety for
disclosure of, inter alia, the various diseases, conditions or
disorders of the CNS relating herein to the first disease or
condition of the present invention, as well as various compounds
and/or therapeutic agents for treating same by application to the
upper 1/3 of the nasal cavity, bypassing of the blood-brain barrier
and subsequent direct delivery of the compounds and/or agents to
the CNS:
U.S. Pat. No. 7,972,595 Methods and compositions for protecting and
treating at least one muscarinic receptor from dysfunction not
resulting from oxidative stress, toxic actions of metals or
infectious agents by administering a pyrophosphate analog;
U.S. Pat. No. 7,786,166 Methods and compositions for protecting and
treating muscarinic receptors through administration of at least
one protective agent;
U.S. Pat. No. 7,776,312 Method of treating Alzheimer's disease
comprising administering deferoxamine (DFO) to the upper one-third
of the nasal cavity;
U.S. Pat. No. 7,618,615 Methods for providing neuroprotection for
the animal central nervous system against neurodegeneration caused
by ischemia;
U.S. Pat. No. 7,084,126 Methods and compositions for enhancing
cellular function through protection of tissue components;
U.S. Pat. No. 6,313,093 Method for Administering Insulin to the
Brain;
US Pat Application 20100061959 Methods for Providing Neuroprotecton
for the Animal Central Nervous System Against the Effects of
Ischemia, Neurodegeneration, Trauma, and Metal Poisoning;
US Patent Application 20080305077 Pharmaceutical Compositions and
Method for Enhancing Targeting of Therapeutic Compounds to the
Central Nervous System;
US Patent Application 20110311654 Methods and Pharmaceutical
Compositions for Treating the Animal Central Nervous System for
Psychiatric Disorders;
US Patent Application 20110236365 Method for Protecting and
Treating at Least One Muscarinic Receptor From Dysfunction
Resulting From Free Radical Damage.
The use of therapeutic agents or compounds that are being used to
treat central nervous system (CNS)-related conditions or diseases
or disorders such as traumatic brain injury (TBI) may cause
unnecessary, unwanted and potentially adverse side effects when
given systemically or by general inhalation methods to the lower
two-thirds of the patient's nasal cavity. In part, this may occur
because systemic uptake dictates that a much larger dose be given,
e.g., orally or intravenously, in order to ensure that an effective
dose actually crosses the blood-brain barrier and enters the CNS.
For example, gastric problems including GI upset, negative effects
on blood pressure, and/or cardiac, liver, or kidney toxicity may
result from systemic administration. Accordingly, a need exists for
a therapeutic agent or compound that may be used to protect the
brain of patients potentially at risk of events that place the
patients at risk of developing TBI. Further, a need exists for a
therapeutic agent or compound that may be used to treat TBI.
Further, a need exists for such a therapeutic agent or compound
that minimizes the adverse side effects generally associated with
administration of drugs used to treat CNS-related disorders. Still
further, a need exists for a delivery system for such a composition
that provides for enhanced uptake of the composition to maximize
the therapeutic affect obtained per administration.
The present invention provides solutions for, inter alia, these
problems.
SUMMARY OF THE INVENTION
The present system is directed in one embodiment to a method of
administration of a therapeutic composition for protecting the
brain of a subject at risk of suffering an injury leading to
traumatic brain injury and/or treatment of injury to the brain
resulting from traumatic brain injury. The method includes
administering one or more therapeutic compositions comprising an
effective amount of insulin directly to the subject patient's CNS,
with no to minimal systemic exposure. Preferably, this method
comprises administration to the upper third of a patient's nasal
cavity, thereby bypassing the patient's blood-brain barrier and
delivering the therapeutic composition directly to the patient's
central nervous system.
BRIEF DESCRIPTION OF THE DRAWINGS
FIG. 1 is a bar graph indicating time for vehicle subjects compared
with insulin subjects to traverse beam walk;
FIG. 2 is a bar graph indicating time for vehicle subjects compared
with insulin subjects to traverse the peg walk;
FIG. 3 is a bar graph indicating expression of CD206 in vehicle
subjects compared with insulin subjects;
FIG. 4 is a bar graph indicating expression of CD86 in vehicle
subjects compared with insulin subjects;
FIG. 5 is a line graph indicating a lack of change in blood glucose
after intranasal insulin delivery (injury-CCI-completed at time=1
h, insulin or saline administered at time=4 h; final blood draw at
time=7 h);
FIG. 6 is a bar graph indicating number of target island crosses
for vehicle subjects compared with insulin subjects in Morris water
maze probe trial;
FIG. 7 is a bar graph indicating search strategy analysis for
vehicle subjects compared with insulin subjects in Morris water
maze probe trial;
FIG. 8a is a line graph illustrating neuronal viability in the
saline rat hippocampus; and
FIG. 8b is a line graph illustrating improved neuronal viability in
the insulin rat hippocampus compared with the saline rat
hippocampus.
DETAILED DESCRIPTION OF THE INVENTION
Definitions
As used herein, "central nervous system" (CNS) refers to the brain
and spinal cord and associated cells and tissues.
As used herein, "systemic administration" refers to administration
of a medication, pharmaceutical and the like by the following
non-limited means: oral, intravenous, intra-arterial,
intramuscular, epidermal, transdermal, subcutaneous, topic,
sublingual as well as general inhalation, i.e., administration to
the lower two-thirds of the patient's nasal cavity. In each of
these cases, the administered drug will migrate through the
patient's circulatory system and, in order to reach the patient's
CNS would be required to cross the patient's blood-brain
barrier.
In the context of the present invention, the terms "treat" and
"therapy" and the like refer to alleviate, slow the progression,
prophylaxis, attenuation or cure of existing disease or condition
that has or is causing cell death in the CNS.
"Protect", as used herein, refers to putting off, delaying,
slowing, inhibiting, or otherwise stopping, reducing or
ameliorating the onset of traumatic brain injury (TBI). It is
preferred that a large enough quantity of the therapeutic agent(s)
and/or compound(s) be applied in non-toxic levels in order to
provide an effective level of activity against TBI in subjects,
e.g., soldiers, police officers, fire fighters, football players
and the like, who are at risk of injury leading to TBI by engaging
in violent activities that may lead to head trauma and TBI or
related concussive injuries. Consequently, patients or subjects
preparing to engage in such violent activities are at risk of
injury leading to TBI and may, therefore, receive protective
effects from the present invention if administered before actually
engaging in said violent activities. Administering an effective
amount of insulin to the patient's brain does, as provided below,
reduce inflammation within the brain. Administering the invention
to the at-risk patient or subject thus provides a
higher-than-normal concentration of insulin within the brain during
the course of the violent activity, once actually engaged in by the
patient or subject. In turn, this concentration of insulin provides
protection against inflammation incurred as a result of head
trauma, working to reduce the inflammation as soon as it appears.
This is in contrast with the patient diagnosed with TBI or other
concussive or inflammatory brain condition. The present invention
may be used to treat the patient's diagnosed TBI, concussive or
inflammatory brain condition.
The method of the present invention may be used with any animal,
such as a mammal or a bird (avian), more preferably a mammal.
Poultry are a preferred bird. Exemplary mammals include, but are
not limited to rats, mice, cats, dogs, horses, cows, sheep, pigs,
and more preferably humans.
An "effective amount" of therapeutic agent(s), i.e., insulin,
and/or component(s) of the pharmaceutical composition of the
present invention comprising therapeutic agent(s) is an amount
sufficient to protect against injury to the brain of a subject at
risk of injury leading to TBI, treat, reduce and/or ameliorate the
symptoms, neuronal damage and/or underlying causes of TBI. In some
instances, an "effective amount" may be sufficient to eliminate the
symptoms of TBI and overcome the disease itself. Preferably, at
least an effective amount of the at least one therapeutic agent,
i.e., insulin, and/or component(s) of the pharmaceutical
composition yields a tissue concentration in the range of about
10.sup.-13 molar to about 10.sup.-9 molar, but the concentrations
may be greater provided that toxicity is avoided. Generally, at
least an effective amount of insulin or pharmaceutical
composition(s) thereof is administered in order to ensure that an
effective amount of insulin is delivered to the target CNS for
protection of the brain of a subject at risk of injury leading to
TBI and treating TBI.
The concentration range of insulin delivered to the upper third of
the patient's nasal cavity may be preferably in the range of
10.sup.-16 molar to about 10.sup.-6 molar in order to yield the
preferable tissue concentration range of about 10.sup.-13 molar to
about 10.sup.-9 molar, though as discussed above, concentrations in
the tissue may be higher so long as toxicity is avoided.
For illustrative purposes only, exemplary treatment regimens
relating generally to the therapeutic agent, i.e., insulin, and/or
pharmaceutical compounds disclosed herein, including dosage ranges,
volumes and frequency are provided below:
Efficacious dosage range for the at least one therapeutic agent,
i.e., insulin and/or vasoconstrictors comprises 1.times.10.sup.-7
to 0.1 mg/kg.
A more preferred dosage range may be 1.times.10.sup.-4 to 0.1
mg/kg.
The most preferred dosage range may be 0.01 to 0.1 mg/kg.
The dosage volume (applicable to nasal sprays or drops) range may
be 0.015 ml-1.0 mi.
The preferred dosage volume (applicable to nasal sprays or drops)
range may be 0.03 ml-0.6 ml.
The brain concentrations that are likely to be achieved with the
dosage ranges provided above are for each of the therapeutic agents
described above, including insulin, for a single dose:
1.times.10.sup.-13 to 1.times.10.sup.-9M.
The present disclosure is generally directed to administering
insulin intranasally to patients for treatment and/or protection of
the brain of a subject at risk of injury leading to TBI of
traumatic brain injury (TBI).
Generally, the method of the present invention comprises protecting
the at-risk brain, or treating TBI with the direct non-invasive
delivery of a therapeutic, i.e., effective, amount or dose of
insulin, or a pharmaceutical composition thereof, to the CNS. This
may be accomplished by administration of at least an effective or
therapeutic amount of insulin, or pharmaceutical composition
thereof, to the upper one-third of the patient's nasal cavity,
thereby delivering the effective or therapeutic amount or dose
directly to the patient's CNS, with minimal systemic exposure.
In some embodiments, the therapeutic agent--insulin--may be
combined with a vasoconstrictor to be administered intranasally to
limit systemic exposure. The vasoconstrictor may be administered to
the nasal cavity prior to administration of the therapeutic
compound to the upper third, or alternatively to the lower
two-thirds, of the nasal cavity or, still more alternatively, the
vasoconstrictor and therapeutic compound may be administered
concurrently, either to the upper one-third or the lower two-thirds
of the patient's nasal cavity. Thus, the present invention allows
for a safe and efficacious treatment, or protection of the at-risk
brain, of a patient's TBI where systemic administration or exposure
is contraindicated.
While not used in conjunction with the treatment of TBI,
administration of intranasal insulin has been shown to improve
memory in both normal adults and in patients with Alzheimer's
disease. Recent studies have shown that insulin may enhance
neuronal activity within the medio-temporal lobe and increase
performance in humans under in-vivo conditions. Impaired insulin
sensitivity may be associated with deficits in verbal fluency and
temporal lobe gray matter volume in the elderly.
There are a variety of types of insulin available that may be used
in accordance with the present disclosure, including insulins for
which zinc is included for stabilization and others which do not
include zinc. Because zinc may be detrimental to the olfactory
system, insulins that do not contain zinc may be preferable in some
cases. Formulations of insulin that either contain no preservatives
(which could be prepared for unit dosing) or a safe preservative
such as pyrophosphate are preferred. In some embodiments the
insulin formulation may not include any phenol or cresol
preservatives.
It is preferred that the neurologic agent--insulin--promote nerve
cell growth and survival or augment the activity of functioning
cells. The neurologic agent may be administered intranasally as a
powder, spray, gel, ointment, infusion, injection, or drops, for
example. The insulin may be administered in an effective dose. The
intranasal composition may be dispensed as a powder or liquid nasal
spray, nose drops, a gel or ointment, through a tube or catheter,
by syringe, by packtail, by pledget, or by submucosal infusion. Any
suitable nasal spray device may be used with embodiments of the
present disclosure.
In some embodiments, the composition may include the neurologic
therapeutic agent (insulin) as well as a vasoconstrictor that may
generally enhance the intranasal therapeutic compound targeting the
CNS, as is further described in U.S. patent application Ser. No.
12/134,385, entitled, "Pharmaceutical Compositions and Methods for
Enhancing Targeting of Therapeutic Compounds to the Central Nervous
System," filed on Jun. 6, 2008, which is hereby incorporated herein
in its entirety. As provided in the aforementioned application,
constriction of blood vessels resulting from action of the
vasoconstrictor in the nasal cavity facilitates transport of the
therapeutic compound(s) or agent(s) into the brain along olfactory
and trigeminal neural pathways, perivascular pathways, or lymphatic
pathways. Thus, intranasal delivery of a therapeutic compound(s) or
agent(s) in combination with an agent that constricts blood vessels
(i.e. a vasoconstrictor) within or in the proximity of the mucosa
of the nasal cavity enhances intranasal drug targeting to, inter
alia, the CNS by reducing absorption into the blood, increasing CNS
concentrations (as well as other targeted locations), or both.
In one embodiment, the pharmaceutical composition may be comprised
of a combination of at least one therapeutic compound comprising
insulin and at least one vasoconstrictor. In another embodiment, at
least one vasoconstrictor may be applied intranasally or otherwise,
i.e., intravenously, topically as a pretreatment or concurrently
with administration of at least one therapeutic compound.
Inclusion of vasoconstrictors in intranasal formulations that
include insulin for protection of the brain of a subject at risk of
injury leading to TBI and/or treatment of TBI may include, but are
not limited to providing the following advantages: reducing
absorption into the blood, which is desirable for drugs with
adverse side effects in the blood or in peripheral tissues;
reducing systemic drug exposure, which is important for drugs that
are rapidly eliminated in drug metabolizing organs or for drugs
that are extensively bound to plasma proteins; targeting drugs to
the olfactory epithelium for CNS delivery of drugs; reducing
clearance of the drug into the blood from the nasal cavity, which
increases the residence time and contact with the nasal epithelium;
targeting drugs to the olfactory epithelium, olfactory bulbs and/or
anterior olfactory nucleus to have therapeutic potential for the
treatment of TBI; targeting high potency drugs to the frontal
cortex to reach brain targets involved in cognition disorders,
motor dysfunction in TBI; and targeting the hippocampus for
treatment of learning and memory disorders associated with TBI.
Exemplary vasoconstrictors in the various embodiments of the
present invention may comprise, without limitation, PHE and/or THZ.
Additional vasoconstrictors will be well known to the skilled
artisan and may include, again without limitation, methoxamine,
phenylephrine, ephedrine, norepinephrine, oxymatazoline,
tetrahydrozoline, xylometazoline, clonidine, guanabenz, guanfacine,
.alpha.-methyldopa, and/or arginine vasopressin.
An at least an effective amount, as herein defined, of the
therapeutic compound, i.e., insulin, and/or vasoconstrictor to be
administered pursuant to embodiments of the invention is the most
preferred method of expression of dosage. Such effective amount is
dependent upon many factors, including but not limited to, the type
of disease or condition giving rise to an anticipated cerebral
ischemia episode, the patient's general health, size, age, and the
nature of the treatment, i.e. short-term or chronic treatment.
Generally, the treatment may be given in a single dose or multiple
administrations, i.e., once, twice, three or more times daily over
a period of time. In some cases, one or more doses daily may be
given over an extended period of time, including, months or
years.
The method of the invention administers an at least an effective
amount of the insulin, or pharmaceutical compound thereof, to the
upper third of the nasal cavity of a mammal. It is preferred that
the at least an effective amount of insulin be administered to the
olfactory area in the upper third of the nasal cavity and
particularly to the olfactory epithelium in order to promote
transport of the agent into the peripheral olfactory neurons rather
than the capillaries within the respiratory epithelium. In some
embodiments it may be preferable to transport insulin to the brain
by means of the nervous system instead of the circulatory system so
that therapeutic agents that are unable to cross the blood-brain
barrier from the bloodstream into the brain may be delivered to
damaged neurons in the brain.
Transportation Pathway to Bypass Blood-Brain Barrier
The Olfactory Nerve
Various methods of the present invention include administration of
at least an effective amount of insulin and/or pharmaceutical
composition(s) thereof to tissue innervated by the olfactory nerve
and that is located in the upper third of the nasal cavity. The at
least an effective amount of insulin and/or pharmaceutical
composition(s) thereof can be delivered to the olfactory area via
application to the upper third of the nasal cavity.
Fibers of the olfactory nerve are unmyelinated axons of olfactory
receptor cells that are located in the upper one-third of the nasal
mucosa. The olfactory receptor cells are bipolar neurons with
swellings covered by hair-like cilia that project into the nasal
cavity. At the other end, axons from these cells collect into
aggregates and enter the cranial cavity at the roof of the nose.
Surrounded by a thin tube of pia, the olfactory nerves cross the
subarachnoid space containing CSF and enter the inferior aspects of
the olfactory bulbs. Once the therapeutic agent(s) and/or
pharmaceutical composition(s) of the present invention is applied
to the upper third of nasal cavity, the therapeutic agent(s) and/or
pharmaceutical composition(s) of the present invention can undergo
transport through the nasal mucosa and into the olfactory bulb and
other areas of the CNS, such as the anterior olfactory nucleus,
frontal cortex, hippocampal formation, amygdaloid nuclei, nucleus
basalis of Meynert, hypothalamus, midbrain, cerebellum, cervical
spinal cord and the like.
Neuronal Transport
Embodiments of the present method includes administration of an at
least an effective amount of insulin and/or pharmaceutical
composition(s) thereof of the present invention to the subject by
application to the upper third of the mammalian subject's nasal
cavity. Application of the at least an effective amount of insulin
and/or pharmaceutical composition(s) thereof of the present
invention in this manner ensures that an effective amount of
insulin and/or pharmaceutical composition(s) are transported to the
CNS, brain, and/or spinal cord along a neural pathway, with reduced
systemic loss and, therefore, minimized systemic exposure. A neural
pathway includes transport within or along a neuron, through or by
way of lymphatics running with a neuron, through or by way of a
perivascular space of a blood vessel running with a neuron or
neural pathway, through or by way of an adventitia of a blood
vessel running with a neuron or neural pathway, or through an
hemangiolymphatic system.
The present invention comprises transportation of the administered
insulin and/or pharmaceutical composition(s) thereof by way of a
neural pathway, rather than through the circulatory system, so that
agent(s) and/or compound(s) that are unable to, or only poorly,
cross the blood-brain barrier from the bloodstream into the brain
can be delivered to the lymphatic system, CNS, brain, and/or spinal
cord. The therapeutic agent(s) and/or pharmaceutical composition(s)
of the present invention, once past the blood-brain barrier and in
the CNS, can then be delivered to various areas of the brain or
spinal cord through lymphatic channels, through a perivascular
space, or transport through or along neurons.
Use of a neural pathway to transport a therapeutic agent(s) and/or
pharmaceutical composition(s) to the brain, spinal cord, or other
components of the central nervous system obviates the obstacle
presented by the blood-brain barrier so that medications, i.e.,
therapeutic agent(s) and/or pharmaceutical compositions of the
present invention, that cannot normally cross that barrier, can be
delivered directly to the CNS, e.g., the brain and spinal cord. In
addition, the present invention can provide for delivery of a more
concentrated level of the therapeutic agent(s) and/or
pharmaceutical composition(s) of the present invention to the CNS
since the therapeutic agent(s) and/or pharmaceutical composition(s)
of the present invention do not become diluted in fluids present in
the bloodstream. As such, the invention provides an improved method
for delivering an effective amount or therapeutic dose of the
administered insulin and/or pharmaceutical composition(s) thereof
directly to the target CNS including the brain and/or spinal
cord.
The Olfactory Neural Pathway
One embodiment of the present method includes delivery of the
effective amount of insulin to the subject's CNS for protection of
the brain of a subject at risk of injury leading to TBI and
treatment of TBI in a manner such that the at least an effective
amount of insulin administered to the upper third of the nasal
cavity is transported into the CNS, e.g., the brain, and/or spinal
cord along an olfactory neural pathway. Typically, such an
embodiment includes administering the at least an effective amount
of insulin and/or other compound(s) to tissue innervated by the
olfactory nerve and inside the nasal cavity. The olfactory neural
pathway innervates primarily the olfactory epithelium in the upper
third of the nasal cavity, as described above. Application of the
at least an effective amount of insulin to a tissue innervated by
the olfactory nerve can deliver an effective amount of insulin
and/or compound(s) to damaged neurons or cells of the CNS,
including but not limited to the brain, and/or spinal cord.
Olfactory neurons innervate this tissue and can provide a direct
connection to the CNS, brain, and/or spinal cord due, it is
believed, to their role in olfaction.
Delivery through the olfactory neural pathway can employ lymphatics
that travel with the olfactory nerve to the various brain areas and
from there into dural lymphatics associated with portions of the
CNS, such as the spinal cord. Transport along the olfactory nerve
can also deliver an effective amount of insulin and/or compound(s)
to an olfactory bulb. A perivascular pathway and/or a
hemangiolymphatic pathway, such as lymphatic channels running
within the adventitia of cerebral blood vessels, can provide an
additional mechanism for transport of an effective amount of
insulin, e.g., to the brain and spinal cord from tissue innervated
by the olfactory nerve.
At least an effective amount of insulin, and/or pharmaceutical
compositions thereof may be administered to the olfactory nerve,
for example, through the olfactory epithelium located at the upper
one-third of the nasal cavity. Such administration can employ
extracellular or intracellular (e.g., transneuronal) anterograde
and retrograde transport of the agent(s) and/or compound(s)
entering through the olfactory nerves to the brain and its
meninges, to the brain stem, or to the spinal cord. Once the at
least an effective amount, i.e., therapeutic dose, of the insulin
and/or pharmaceutical composition thereof is dispensed into or onto
tissue innervated by the olfactory nerve, the administered insulin
and/or pharmaceutical composition and/or components thereof may be
transported through the tissue and travel along olfactory neurons
into areas of the CNS including but not limited to the brain stem,
cerebellum, spinal cord, cerebrospinal fluid, olfactory bulb, and
cortical and subcortical structures. Thus, an effective amount of
insulin and/or pharmaceutical composition thereof, is delivered to
the target CNS for protection of the brain of a subject at risk of
injury leading to TBI and/or treatment of TBI.
The blood-brain barrier is bypassed in the present invention by
application of at least an effective amount of insulin and/or
pharmaceutical composition(s) comprising insulin and/or
composition(s) or compound(s) to the upper third of the nasal
cavity of the patient, e.g., a mammal. The administered amount of
the insulin and/or pharmaceutical composition thereof of the
invention migrate from the nasal mucosa through foramina in the
cribriform plate along the olfactory neural pathway and an
effective amount is delivered directly into the CNS. Further,
vasoconstrictors may be applied to the nasal cavity of the patient,
either before or during the application of the at least an
effective amount of insulin and/or pharmaceutical composition(s)
thereof to the upper third of the patient's nasal cavity, to
enhance the efficiency of delivery of the an effective amount of
insulin to the patient's CNS and minimization of any potential
systemic exposure of the administered insulin.
Administration to the nasal cavity employing a neural pathway can
thus deliver an effective amount of therapeutic agent(s), e.g.,
insulin and/or pharmaceutical compositions thereof to the lymphatic
system, brain stem, cerebellum, spinal cord, and cortical and
subcortical structures of the mammalian patient. The therapeutic
agent(s), e.g., insulin and/or pharmaceutical composition(s)
thereof of the present invention alone may facilitate this movement
into the CNS, i.e., brain, and/or spinal cord. Alternatively, a
carrier may assist in the transport of the administered insulin
and/or pharmaceutical composition of the present invention into and
along the neural pathway. Administration of the insulin and/or
pharmaceutical composition(s) thereof of the present invention to
the upper third of the mammalian patient's nasal cavity thus
enables bypassing of the blood-brain barrier through a transport
system from the nasal mucosa and/or epithelium to the CNS, i.e.,
brain and spinal cord where an effective amount of the administered
insulin is delivered.
Various embodiments of the invention administer an at least an
effective amount of insulin and/or pharmaceutical composition(s)
thereof of the present invention to tissue innervated by the
olfactory nerves. Such nerve systems can provide a direct
connection between the outside environment and the brain, thus
providing advantageous delivery of the agent(s) and/or compound(s)
to the CNS, including brain, brain stem, and/or spinal cord. The
administered insulin and/or pharmaceutical composition(s) thereof
of the present invention may be unable to cross or inefficiently
cross the blood-brain barrier from the bloodstream into the brain.
Alternatively, for those agent(s) and/or composition(s) that may
cross the blood-brain barrier, the present invention offers an
alternative treatment for those patients having a concurrent
system, non-CNS disease or condition that contraindicates systemic
administration of the therapeutic agent(s) and/or compositions(s)
needed within the CNS to treat a first CNS-related disease,
condition or disorder. Thus, the methods of the present invention
allow for the delivery of an effective amount of insulin and/or
pharmaceutical composition(s) thereof to the target CNS by way of
the olfactory nerve rather than through the circulatory system in
order to facilitate protection of the brain of a subject at risk of
injury leading to TBI and/or treatment of TBI. Thus, this method of
administration of at least an effective amount of insulin to the
upper third of the nasal cavity and delivery of the effective
amount of insulin to the target CNS allows for the efficient and
non-invasive delivery of an effective amount of insulin and/or
pharmaceutical composition(s) thereof of the present invention to
the CNS, brain, or spinal cord without systemic loss or
exposure.
Alternative Pathways
Alternative non-systemic pathways to the olfactory nerve pathway
discussed above comprise pathways along other nerves that innervate
the nasal cavity, e.g., the trigeminal pathway, well known to the
skilled artisan.
Administration of Therapeutic Agents) and/or Pharmaceutical
Compounds
Administering insulin according to the methods of the invention for
protection of the brain of a subject at risk of injury leading to
TBI and/or treatment of TBI may include application of at least an
effective amount of the therapeutic agent, i.e., insulin alone or
formulating the at least an effective amount of insulin with at
least an effective amount of one or more of the compounds described
supra as pharmaceutical compositions and administering the
pharmaceutical compositions to a mammalian subject or host,
including a human patient, intranasally to the upper third of the
nasal cavity. The therapeutic agent(s) and/other components of the
pharmaceutical composition thereof, e.g., vasoconstrictor may be
administered at one of a variety of doses sufficient to provide an
effective amount at the desired point of action in the CNS for the
administered at least an effective amount of insulin and/or
pharmaceutical composition component.
As noted, vasoconstrictor(s) may be delivered as pre-treatment,
co-treatment and/or post-treatment with the therapeutic agent(s)
and/or pharmaceutical composition, either alone or as a component
of the pharmaceutical composition. Delivery of at least an
effective amount of insulin in this manner results in delivery of
an effective amount of insulin to the target CNS with maximum
efficiency in the delivery of insulin, i.e., with minimal to no
systemic exposure of insulin.
For application to the upper third of the nasal cavity as
suspensions, aerosols, sprays or drops, the at least an effective
amount of insulin and/or pharmaceutical composition(s) can be
prepared according to techniques well known in the art of
pharmaceutical formulation. The compositions can be prepared as
suspensions of the agent(s) in solutions which may comprise salts
such as saline, components such as phosphate, succinate or citrate
buffers to maintain pH, osmoregulatory and osmotic agents such as
taurine, and suitable preservatives, absorption promoters to
enhance bioavailability, fluorocarbons or other solubilizing or
dispersing agents known in the art. The means of applying a
pharmaceutical composition intranasally to the upper third of the
nasal cavity may be in a variety of forms such as a powder, spray,
gel or nose drops.
Other forms of compositions for administration of the at least an
effective amount of insulin and/or pharmaceutical compositions or
elements thereof include a suspension of a particulate, such as an
emulsion, a liposome, or in a sustained-release form to prolong the
presence of the pharmaceutically active agent in an individual. The
powder or granular forms of the pharmaceutical composition may be
combined with a solution and with a diluting, dispersing or
surface-active agent. Additional compositions for administration
include a bioadhesive to retain the agent at the site of
administration at the upper third of the nasal cavity, for example
a spray, paint, or swab applied to the mucosa. A bioadhesive can
refer to hydrophilic polymers, natural or synthetic, which, by the
hydrophilic designation, can be either water soluble or swellable
and which are compatible with the pharmaceutical composition. Such
adhesives function for adhering the formulations to the mucosal
tissues of the upper third of the nasal cavity. Such adhesives can
include, but are not limited to, hydroxypropyl cellulose,
hydroxypropyl methylcellulose, hydroxy ethylcellulose,
ethylcellulose, carboxymethyl cellulose, dextran, gaur gum,
polyvinyl pyrrolidone, pectins, starches, gelatin, casein, acrylic
acid polymers, polymers of acrylic acid esters, acrylic acid
copolymers, vinyl polymers, vinyl copolymers, polymers of vinyl
alcohols, alkoxy polymers, polyethylene oxide polymers, polyethers,
and combinations thereof. The composition can also be in the form
of lyophilized powder, which can be converted into solution,
suspension, or emulsion before administration. The pharmaceutical
composition is preferably sterilized by membrane filtration and is
stored in unit-dose or multi-dose containers such as sealed vials
or ampoules.
The pharmaceutical composition may be formulated in a
sustained-release form to prolong the presence of the active
therapeutic agent(s) in the treated individual. Many methods of
preparation of a sustained-release formulation are known in the art
and are disclosed in Remington's Pharmaceutical Sciences.
Generally, the therapeutic agent(s), pharmaceutical composition
and/or components of the pharmaceutical composition, i.e.,
vasoconstrictor may be entrapped in semi-permeable matrices of
solid hydrophobic polymers. The matrices can be shaped into films
or microcapsules. Matrices can include, but are not limited to,
polyesters, copolymers of L-glutamic acid and gamma
ethyl-L-glutamate, polylactides, polylactate polyglycolate,
hydrogels, non-degradable ethylene-vinyl acetate, degradable lactic
acid-glycolic acid copolymers, and alginic acid suspensions.
Suitable microcapsules can also include hydroxymethylcellulose or
gelatin and poly-methyl methacrylate. Microemulsions or colloidal
drug delivery systems such as liposomes and albumin microspheres
can also be used.
Delivery Systems
The therapeutic agent, i.e., insulin, and/or a pharmaceutical
composition comprising at least an effective dose of insulin and/or
components of the pharmaceutical composition of the present
invention may further be dispensed and applied to the upper third
of the nasal cavity as a powdered or liquid nasal spray,
suspension, nose drops, a gel, film or ointment, through a tube or
catheter, by syringe, by packtail, by pledget (a small flat
absorbent pad), by nasal tampon or by submucosal infusion. In some
aspects of the present invention, the methods comprise
administering to an individual the at least an effective dose of
insulin and/or a pharmaceutical composition thereof to the upper
third of the nasal cavity by way of a delivery device. Nasal drug
delivery can be carried out using devices including, but not
limited to, unit dose containers, pump sprays, droppers, squeeze
bottles, airless and preservative-free sprays, nebulizers (devices
used to change liquid medication to an aerosol particulate form),
metered dose inhalers, and pressurized metered dose inhalers. In
some aspects, an accurate effective dosage amount is contained
within a bioadhesive patch that is placed directly within and on
the upper third of a nasal cavity.
At least an effective dose of insulin and/or a pharmaceutical
composition comprising at least an effective dose of insulin and/or
components of the therapeutic composition of the present invention
may be conveniently delivered to the upper third of the nasal
cavity in the form of an aerosol spray using a pressurized pack or
a nebulizer and a suitable propellant including, but not limited
to, dichlorodifluoromethane, trichlorofluoromethane,
dichlorotetrafluoroethane, hydrocarbons, compressed air, nitrogen
or carbon dioxide. An aerosol system requires the propellant to be
inert towards the therapeutic agent(s) and/or pharmaceutical
composition as will be readily recognized by the skilled artisan.
In the case of a pressurized aerosol, the dosage unit may be
controlled by providing a valve to deliver an accurately metered
amount.
The means to deliver the at least an effective amount of insulin or
pharmaceutical composition comprising the at least an effective
amount of insulin and/or components of the pharmaceutical
composition of the present invention to the upper third of the
nasal cavity as a powder may be in a form such as microspheres
delivered by a nasal insufflator device (a device to blow a gas,
powder, or vapor into a cavity of the body) or pressurized aerosol
canister. The insufflator produces a finely divided cloud of the
dry powder or microspheres. The insufflator may be provided with
means to ensure administration of a substantially metered amount of
the pharmaceutical composition. The powder or microspheres should
be administered in a dry, air-dispensable form. The powder or
microspheres may be used directly with an insufflator which is
provided with a bottle or container for the powder or microspheres.
Alternatively the powder or microspheres may be filled into a
capsule such as a gelatin capsule, or other single dose device
adapted for nasal administration. The insufflator can have means
such as a needle to break open the capsule or other device to
provide holes through which jets of the powdery composition can be
delivered to the upper third of the nasal cavity.
Intermittent and Cyclic Dosing
In various embodiments of the invention, therapeutic agent, i.e.,
insulin, and/or a pharmaceutical composition comprising at least an
effective amount of insulin and/or the components of the
pharmaceutical composition may be administered as a single and
one-time dose, or alternatively the at least an effective amount of
insulin and/or the components of the pharmaceutical composition may
be administered more than once and intermittently. By "intermittent
administration" is intended administration of at least an effective
amount of insulin and/or the components of the pharmaceutical
composition, followed by a time period of discontinuance, which is
then followed by another administration of the at least effective
amount, and so forth. Administration of the at least an effective
amount of insulin and/or the components of the pharmaceutical
composition may be achieved in a continuous manner, as for example
with a sustained-release formulation, or it may be achieved
according to a desired daily dosage regimen, as for example with
one, two, three, or more administrations per day. By "time period
of discontinuance" is intended a discontinuing of the continuous
sustained-released or daily administration of the insulin and/or
the components of the pharmaceutical composition. The time period
of discontinuance may be longer or shorter than the period of
continuous sustained-release or daily administration. During the
time period of discontinuance, the concentration(s) of insulin
and/or the components of the pharmaceutical composition level in
the relevant tissue is substantially below the maximum level
obtained during the treatment. The preferred length of the
discontinuance period depends on the concentration of the effective
dose and the form of therapeutic agent(s) and/or the components of
the pharmaceutical composition used. The discontinuance period can
be at least 2 days, preferably is at least 4 days, more preferably
is at least 1 week and generally does not exceed a period of 4
weeks. When a sustained-release formulation is used, the
discontinuance period must be extended to account for the greater
residence time of the at least one therapeutic agent at the site of
injury. Alternatively, the frequency of administration of the
effective dose of the sustained-release formulation can be
decreased accordingly. An intermittent schedule of administration
of insulin and/or the components of the pharmaceutical composition
may continue until the desired therapeutic effect, and ultimately
treatment of the disease or disorder is achieved.
In yet another embodiment, intermittent administration of the at
least an effective amount(s) of insulin and/or the components of
the pharmaceutical composition is cyclic. By "cyclic" is intended
intermittent administration accompanied by breaks in the
administration, with cycles ranging from about 1 month to about 2,
3, 4, 5, or 6 months. For example, the administration schedule
might be intermittent administration of the at least an effective
dose of insulin and/or the components of the pharmaceutical
composition, wherein a single short-term dose is given once per
week for 4 weeks, followed by a break in intermittent
administration for a period of 3 months, followed by intermittent
administration by administration of a single short-term dose given
once per week for 4 weeks, followed by a break in intermittent
administration for a period of 3 months, and so forth. As another
example, a single short-term dose may be given once per week for 2
weeks, followed by a break in intermittent administration for a
period of 1 month, followed by a single short-term dose given once
per week for 2 weeks, followed by a break in intermittent
administration for a period of 1 month, and so forth. A cyclic
intermittent schedule of administration of insulin and/or the
components of the pharmaceutical composition to a subject may
continue until the desired therapeutic effect, and ultimately
treatment of the disorder or disease is achieved.
Working Example
In a reduction to practice of one embodiment of the present
invention, a Moderate Controlled Cortical Impact Injury Model,
developed for use in establishing a Traumatic Brain Injury (TBI)
model was employed. In this study, adult male Sprague Dawley rats
were subjected to a controlled cortical impact as that procedure is
described in "A mouse model of sensorimotor controlled cortical
impact: characterization using longitudinal magnetic resonance
imaging, behavioral assessments and histology". Onyszchuk G,
Al-Hafez B, He Y Y, Bilgen M, Berman N E, Brooks W M. J Neurosci
Methods. 2007 Mar. 15; 160(2):187-96. Onyszchuk, G., et al., J.
Neurosci. Methods, 2007, 160(2): p. 187-196, the contents of which
are hereby incorporated by reference.
Procedure
Following the controlled injury using the TBI model described
above, the injured rats received either saline or insulin under
anesthesia by isoflurane. A first dose of either saline (saline
rats) or insulin (insulin rats) was delivered 4 hours post-injury
to the upper third of the nasal cavity as described above in
connection with the present invention. Thereafter, doses of saline
or insulin were administered once a day for 7 to 14 days to the
upper third of the nasal cavity. Motor function was evaluated in
the saline rats and with the insulin rats. In addition,
immunohistochemistry and Western Blot analyses were performed with
a focus on inflammatory cells, microglia, and neuronal
survival.
Results
Blood was taken from the tail vein of a small cohort of rats (n=3
per treatment group) immediately prior to injury, 4 hours after
injury/immediately before treatment and 3 hours after treatment.
The results indicate no significant difference in blood glucose
between the two groups, shown in FIG. 5. Consequently, the
administration of insulin to the upper third of the nasal passage
does not significantly affect the patient's blood glucose.
In addition, both the saline rats and the insulin rats were weighed
on days 1, 7, 14 and 21 post-injury. The results indicate no
significant difference in weight between the two groups.
Consequently, the administration of insulin to the upper third of
the nasal passage does not significantly affect the patient's
weight.
Motor function tests consisting of a beam walk and a peg walk, both
tests well-established measures of functional deficit and recovery
in preclinical TBI research, also given to the saline rats and the
insulin rats on days 1 and 7 post-injury. The results for the beam
walk, shown in FIG. 1, indicate a significant decrease in time for
the insulin rats to cross the beam as compared with day 1 baseline
results and as compared with the saline (vehicle) rats performance
on day 7.
The peg walk results illustrated in FIG. 2 indicate a positive
trend towards improved motor function in the insulin rat group from
baseline results from day 1 as well as when compared with saline
(vehicle) rats on day 7. Thus, intranasal insulin, administered
according to the present invention to the upper third of the nasal
cavity in subjects with TBI results in improved motor function in
those subjects.
Blood glucose levels were also tested following intranasal insulin
delivery. FIG. 5 illustrates a lack of change in blood glucose
after intranasal insulin delivery (injury-CCI-completed at time=1
h, insulin or saline administered at time=4 h; final blood draw at
time=7 h).
Cognitive function was also tested in saline (vehicle) and insulin
treated rats on days 11-14 post-injury, using the well-known and
well-justified Morris water maze task. Memory retention was
assessed using the probe trial portion of the task, and showed that
insulin-treated rats had a significant increase in the number of
crosses of the target island region in comparison to saline
(vehicle) treated rats, where they had been previously trained to
exit the maze, as shown in FIG. 6. In addition, search strategy,
which indicates method of searching the maze for the exit island,
shown in FIG. 7, demonstrates that saline (vehicle) treated rats
utilized significantly less targeted search methods (looping) than
insulin rats.
In addition to the above motor and cognitive function tests, the
effect of insulin administered to the upper third of the nasal
cavity in the insulin rat group as compared with the saline rat
group was also evaluated by using an immunohistochemical marker of
neurons, specifically NeuN, to examine the effect of the insulin on
neurons in the hippocampus post-injury after sacrifice on day 8.
FIGS. 8a and 9b are photographs that indicate, following a
quantitative assessment according to well-known methods, an
improved neuronal viability in the hippocampus of the intranasal
rats as compared with the saline rats. FIG. 8a illustrates neuronal
viability in the saline rat hippocampus. FIG. 8b illustrates an
improved neuronal viability in the insulin rat hippocampus, as
illustrated by the brighter and more robust neuronal presence as
compared with the saline rat of FIG. 8a.
Finally, the effect of insulin administered as in the Working
Example on microglia was evaluated post-injury and following
sacrifice on day 8 for the saline rats and the insulin rats.
Microglia are the macrophages of the brain, with two dominant
phenotypes: M1 and M2.
M1 microglia are classically activated pro-inflammatory cells. M1
microglia are useful in the initial healing process, but persistent
activation results in neuronal cell death due to production of
reactive oxygen species. M2 microglia are anti-inflammatory,
pro-healing macrophages.
The Working Example results, illustrated in FIGS. 3 and 4 indicate
a significant increase in M2 microglia, characterized by expression
of CD206, in the intranasal rats as compared with the intranasal
saline rats as shown in FIG. 3. There was no significant increase
in expression of M1 microglia markers, characterized by expression
of CD86 and as illustrated in FIG. 4. This indicates, inter alia,
that the insulin regimen according to the present invention and as
used in the Working Example is pushing the microglia toward the M2
anti-inflammatory phenotype. Thus, subjects with TBI that are
treated with insulin administered to the upper third of the
subject's nasal cavity experience an increase in activation of
anti-inflammatory cells, specifically M2 microglia cells which, in
turn promote healing of the damaged cells in the subject with TBI,
or protect against damage to the brain of a subject at risk of
injury to the brain leading to TBI.
Further, inflammation is a major, and damaging, component of
neurodegenerative central nervous system disorders, including but
not limited to Alzheimer's disease, Parkinson's disease, ALS,
Huntington's disease, to name a few. As a result, the discovery
that intranasal insulin increases the M2 anti-inflammatory
microglia phenotype in the brain of a patient experiencing
neuroinflammation in the brain as a result of brain injury or other
neurodegenerative disorder, condition and/or disease is applicable
and useful to protect, prevent and/or treat such inflammation.
CONCLUSION
The Working Example indicates that insulin administered to the
upper third of the patient's nasal cavity, increases neuronal
viability in the hippocampus, increased the formation of
anti-inflammatory M2 microglia which promote healing, and increased
motor and cognitive functional recovery after Traumatic Brain
Injury (TBI).
In addition, the results of the Working Example indicate that
insulin delivered to the brain of patients with brain injury or
other neurodegenerative disorder involving inflammation of the
brain can be used to reduce the associated inflammation by
increasing production of the M2 anti-inflammatory phenotype in the
patient's brain. Certainly, neurodegenerative disorders such as
Alzheimer's disease, Parkinson's disease, Huntington's disease, ALS
involve inflammation of the brain and are, therefore, amenable to
treatment using the present invention. In addition, patients at
risk of developing a neurodegenerative condition and/or disease
leading to neuroinflammation may be identified and the
neuroinflammation prevented, or protected against.
The invention has been described with reference to various specific
and preferred embodiments and techniques. However, it should be
understood that many variations and modifications may be made while
remaining within the spirit and scope of the invention.
* * * * *
References